摘要目的 建立通用型荧光探针用于不同成熟型微RNA的定量检测.方法 基于茎环法发明者发明且目前应用最为广泛的茎环框架,采用Primer Premier 5.0软件在其茎环框架内部设计了一通用的荧光探针及不同的引物对.采用以上探针和引物对对hsa-miR-143-3p,hsa-miR-145-5p,hsa-miR-124-3p,hsa-miR-339-5p,hsa-miR-361-5p,hsa-miR-423-3p和hsa-miR-486-5p进行定量检测.结果 结果表明该通用型荧光探针可用于不同成熟型微RNA(hsa-miR-143-3p,hsa-miR-145-5p,hsa-miR-124-3p,hsa-miR-339-5p,hsa-miR-361-5p,hsa-miR-423-3p,hsa-miR-486-5p)的定量检测.结论该通用型探针可用于茎环法定量检微RNA,可让定量更加准确、实验成本更低.理论上该探针可以用于所有的成熟型微RNA进行荧光定量PCR检测.

abstractsObjective To establish a universal probe for quantitative detection of different mature microRNAs. Methods Based on the most widely used stem-loop framework which was invented by the inventors of stem-loop method,Primer Premier 5.0 software was used to design a universal fluorescent probe and different primer pairs in the stem-loop framework. The probe and the primer pairs were used for quantitative detection of hsa-miR-333-5p,hsa-miR-361-5p,hsa-miR-333-5p,hsa-miR-361-5p,hsa-miR-MiR-423-3p and hsa-miR-486-5p. Results The findings indicated that the universal fluorescent probe could be used for quantitative detection of different mature microRNAs(hsa-miR-143-3p,hsa-miR-145-5p,hsa-miR-124-3p,hsa-miR-339-5p,hsa-miR-361-5p,hsa-miR-423-3p,and hsa-miR-486-5p). Conclusion The universal probe can be used for quantitative detection of RNAi by stem loop method,which can make the quantitative more accurate with lower experimental cost. Theoretically,the probe can be used for fluorescent quantitative PCR detection of all mature microRNAs.